Digest mix

Procedure

Vortex all reagents before use.

Add appropriate amount of deionized H2O to sterile 0.6 mL tube

Add restriction enzyme buffer.

Add BSA.

Add DNA.

Add each enzyme. Also, the enzyme is in some percentage of glycerol which tends to stick to the sides of your tip. To ensure you add only 1 μL, just touch your tip to the surface of the liquid when pipetting.

Incubate for 2 hours at 37°C.

Incubate for 20 mins at 80°C to heat inactivate enzyme. This step is sufficient to inactivate even Pst I.